Seminal fluid factors regulate activin A and follistatin synthesis in female cervical epithelial cells

Sharkey, David J.; Schjenken, John E.; Mottershead, David G.; Robertson, Sarah A.

Abstract

Background:
Overexpression of microRNA-31 (miR-31) is implicated in the pathogenesis of esophageal squamous cell carcinoma (ESCC), a deadly disease associated with dietary zinc deficiency. Using a rat model that recapitulates features of human ESCC, the mechanism whereby Zn regulates miR-31 expression to promote ESCC is examined.

Methods:
To inhibit in vivo esophageal miR-31 overexpression in Zn-deficient rats (n = 12–20 per group), locked nucleic acid–modified anti-miR-31 oligonucleotides were administered over five weeks. miR-31 expression was determined by northern blotting, quantitative polymerase chain reaction, and in situ hybridization. Physiological miR-31 targets were identified by microarray analysis and verified by luciferase reporter assay. Cellular proliferation, apoptosis, and expression of inflammation genes were determined by immunoblotting, caspase assays, and immunohistochemistry. The miR-31 promoter in Zn-deficient esophagus was identified by ChIP-seq using an antibody for histone mark H3K4me3. Data were analyzed with t test and analysis of variance. All statistical tests were two-sided.

Results:
In vivo, anti-miR-31 reduced miR-31 overexpression (P = .002) and suppressed the esophageal preneoplasia in Zn-deficient rats. At the same time, the miR-31 target Stk40 was derepressed, thereby inhibiting the STK40-NF-κΒ–controlled inflammatory pathway, with resultant decreased cellular proliferation and activated apoptosis (caspase 3/7 activities, fold change = 10.7, P = .005). This same connection between miR-31 overexpression and STK40/NF-κΒ expression was also documented in human ESCC cell lines. In Zn-deficient esophagus, the miR-31 promoter region and NF-κΒ binding site were activated. Zn replenishment restored the regulation of this genomic region and a normal esophageal phenotype.

Conclusions:
The data define the in vivo signaling pathway underlying interaction of Zn deficiency and miR-31 overexpression in esophageal neoplasia and provide a mechanistic rationale for miR-31 as a therapeutic target for ESCC.

Citation

Sharkey, D. J., Schjenken, J. E., Mottershead, D. G., & Robertson, S. A. (2015). Seminal fluid factors regulate activin A and follistatin synthesis in female cervical epithelial cells. Molecular and Cellular Endocrinology, 417(11), 178-190. https://doi.org/10.1016/j.mce.2015.09.022

• Journal Article Type: Article
• Publication Date: 2015-12
• Deposit Date: Jul 4, 2023
• Journal: Molecular and Cellular Endocrinology
• Print ISSN: 0303-7207
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 417
• Issue: 11
• Pages: 178-190
• DOI: https://doi.org/10.1016/j.mce.2015.09.022
• Keywords: Endocrinology; Molecular Biology; Biochemistry
• Additional Information: This article is maintained by: Elsevier; Article Title: Seminal fluid factors regulate activin A and follistatin synthesis in female cervical epithelial cells; Journal Title: Molecular and Cellular Endocrinology; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.mce.2015.09.022; Content Type: article; Copyright: Crown copyright © 2015 Published by Elsevier Ireland Ltd. All rights reserved.